TABLE 1

Sequences are shown below for one of two complementary oligonucleotides used in the construction of the indicated mutation(s) Bold indicates changes from the wild-type sequence. Underline indicates the location of the desired mutation. Italics indicate additional synonymous changes that alter a restriction site and were used to facilitate identification of plasmids containing the desired nonsynonymous mutation. Multiple mutations were made by sequential addition of changes using these same oligonucleotides.


CYP2A6 Mutation

Sequence (5′ to 3′)

Restriction Site Altered
V110L GCC ACC TTC GAC TGG CTG TTT AAA GGC TAT GGC Add DraI
V117A GGC TAT GGC GTG GCA TTC AGC AAC GGG Add BbsI
I208S GC ATG ATG CTA GGA AGC TTC CAG TTC ACG TCA ACC TCC Delete HindIII
S213A GGA ATC TTC CAG TTC ACG GCG ACG TCC ACG GGG CAG C Add AatII
I300F CG TTG AAC CTC TTC ATC GGG GGT ACT GAG ACC GTG AGC Add KpnI
G301A CG TTG AAC CTC TTC ATT GCG GGT ACC GAG ACC GTC AGC Add KpnI
V365M CAA AGA TTT GGA GAC ATG ATC CCC ATG AGT TTG None
I366L C CAA AGA TTT GGA GAC GTC CTG CCC ATG AGT TTG GCC C Add AatII
S369G C GTG ATC CCC ATG GGT TTG GCC CGC AGA GTC AAA AAG G Delete NcoI
R372H CCC ATG AGT TTG GCC CAC AGA GTG AAA AAG GAC ACC Delete DraIII
I300F/G301A CC ACG TTG AAC CTC TTC TTC GCG GGT ACC GAG ACC GTC AGC ACC Add KpnI
V365M/I366L
CAA AGA TTT GGA GAC ATG CTG CCC ATG AGT TTG GCC CGC
None